Background: Respiratory viral and bacterial infections are one of the leading causes of hospitalization and death worldwide, particularly affecting children, the elderly, and immune compromised individuals. The detection of causative pathogens is crucial for understanding the epidemiology, prevention, management, and treatment of severe pneumonia. Molecular diagnostic methods provide high sensitivity and speed, enabling the simultaneous detection of viral and bacterial co-infections. Aim: Detection of single and combined viral and bacterial infections in hospitalized children with pneumonia using multiplex PCR. Materials and Methods: We conducted a cross-sectional study design involving 100 hospitalized participants at the National Center for Mothers, Newborns, and Women II. The study utilized questionnaires and laboratory analysis methods. Nasopharyngeal swabs were collected from the participants using a sterile swab and transported in a UTM transport medium (Jiangsu, China). After nucleic acid extraction using the STARMag 96 ProPrepC assay, the samples were analyzed with the Seeprep 32 automated analyzer (Seegene Inc., Korea). Real-time polymerase chain reaction (PCR) analysis was performed using the Allplex™ RV Master Assay and Allplex™ PneumoBacter Assay (Seegene Inc., Korea), with the CFX-96 system (BioRad Inc., USA). Results: The prevalence of viral infections in the study population was as follows: influenza type A 3% (n=3), influenza type B 3% (n=3), total influenza virus 14% (n=14), respiratory syncytial virus 18% (n=18), adenovirus 10% (n=10), rhinovirus 20% (n=20), and SARS-CoV-2 5% (n=5). No bacterial infections, such as B.parapertussis or L.pneumophila, were detected. However, bacterial infections identified included B.pertussis in 1% (n=1), C.pneumoniae in 2% (n=2), H.influenzae in 43% (n=43), M.pneumoniae in 52% (n=52), and S.pneumoniae in 45% (n=45). Of the 100 children in the study, 6% had influenza or influenza-like illness. Among these, 43% had the virus alone, and 14% had a viral co-infection. Bacterial infections were detected individually in 40% of cases, with co-infections in 46% of bacterial cases. Conclusion Metapneumovirus, B.parapertussis, and Legionella pneumophila were not detected in the study participants (0%). Among the viral infections, respiratory syncytial virus (18%, n=18) and rhinovirus (20%, n=20) had the highest prevalence. For bacterial infections, H.influenzae (43%, n=43), M.pneumoniae (52%, n=52), and S.pneumoniae (45%, n=45) were most commonly detected.